Exclusion of Sall 4 as the sex-determining gene in the Mandarin vole Microtus mandarinus mandarinus

In previous studies, we have shown that the Sry HMG-box is absent in Microtus mandarinus mandarinus (M. m. mandarinus), suggesting that sex determination of M. m. mandarinus is independent of the Sry gene. We amplified a 312 bp fragment within exon 2 of the Sall4 gene in the mouse and M. m. mandarinus using polymerase chain reaction (PCR) and detected Sall4 using fluorescence in situ hybridization (FISH) technique. The probes for the Sall4 gene were labeled with digoxigenin using PCR and hybridized to chromosomes and interphase nuclei of the mouse and M. m. mandarinus. Our results suggested that Sall4 exists in the genome of male and female M. m. mandarinus, and the sequence within exon 2 of the gene is the same in the mouse and M. m. mandarinus. The results also showed that Sall4 is localized on chromosome 6 in M. m. mandarinus. As they are the sex chromosomes in M. m. mandarinus, the results excluded the Sall4 gene from being the testis-determining factor in this species. We propose that in M. m. mandarinus, sex determination is controlled by another yet unknown gene on the sex chromosomes.     

Geomorphometric differences among four species of Microtus in Turkey (Mammalia: Rodentia)

In order to determine the phenotypic associations of four morphologically similar species of Microtus occurring in Turkey, we applied landmark-based shape analysis. The skulls of Microtus anatolicus, M. dogramacii, M. guentheri and M. levis (= M. rossiaemeridionalis) were found to differ significantly in terms of both size and shape. M. guentheri had the biggest skull, while M. levis had the smallest. Sexual dimorphism was found in the shape of the skull in M. dogramacii and M. levis. The tympanic bulla area is enlarged in M. anatolicus compared to the other species. Mahalanobis distances (the distance between a point and the group mean, taking into account the within-group covariance-variance matrix) confirm the distinction of the arvalis group (M. levis) and socialis group (Microtus anatolicus, M. dogramacii, M. guentheri).     

A Novel MS7 Repeat Specific for Heterochromatin of Sex Chromosomes in Voles of Genus Microtus, Group arvalis: Genomic Organization and Chromosomal Localization

The tandemly arranged MS4 repeat with monomeric units of 4.1 kb is species-specifically distributed in heterochromatin of sex chromosomes of four common vole species of genus Microtus, group arvalis. In this work, we studied the genomic organization of the MS4 homolog in euchromatin of the X chromosome of M. arvalis. It has been shown by analyzing the phage genomic clones that one MS4 copy makes a part of a monomeric unit exceeding 8.5 kb that also includes a new MS7 repeat and, possibly, LINE fragments. MS7 is located together with MS4 in heterochromatin of common vole sex chromosomes, but in a substantially lesser amount. Probably, as a result of an evolutionary transition of an original repeat from euchromatin of the X chromosome to heterochromatin of the Y chromosome, MS4 underwent multiple amplification, and MS7 spread throughout heterochromatin, being surrounded by the MS4 tandem arrays.     

Dental Variation in Sibling Species <Emphasis Type="Italic">Microtus arvalis</Emphasis> and <Emphasis Type="Italic">M. rossiaemeridionalis</Emphasis> (Arvicolinae,Rodentia): Between-Species Comparisons and Geography of Morphotype Dental Patterns

The data on dental variability in natural populations of sibling species of common voles (“arvalis” group, genus Microtus) from European and Asian parts of the species’ ranges are summarized using a morphotype-based approach to analysis of dentition. Frequency distributions of the first lower (m1) and the third upper (M3) molar morphotypes are analyzed in about 65 samples of M. rossiaemeridionalis and M. arvalis represented by arvalis and obscurus karyotypic forms. Because of extreme similarity of morphotype dental patterns in the taxa studied, it is impossible to use molar morphotype frequencies for species identification. However, a morphotype-based approach to analysis of dental variability does allow analysis of inter-species comparisons from an evolutionary standpoint. Three patterns of dental complexity are established in the taxa studied: simple, basic (the most typical within the ranges of both species), and complex. In M. rossiaemeridionalis and in M. arvalis obscurus only the basic pattern of dentition occurs. In M. arvalis arvalis, both simple and basic dental patterns are found. Analysis of association of morphotype dental patterns with geographical and environmental variables reveals an increase in the number of complex molars with longitude and latitude: in M. arvalis the pattern of molar complication is more strongly related to longitude, and in M. rossiaemeridionalis—to latitude. Significant decrease in incidence of simple molars with climate continentality and increasing aridity is found in M. arvalis. The simple pattern of dentition is found in M. arvalis arvalis in Spain, along the Atlantic coast of France and on islands thereabout, in northeastern Germany and Kirov region in European Russia. Hypotheses to explain the distribution of populations with different dental patterns within the range of M. arvalis sensu stricto are discussed.     

Local coexistence and niche differences between the Lusitanian and Mediterranean pine voles (<Emphasis Type="Italic">Microtus lusitanicus</Emphasis> and <Emphasis Type="Italic">M. duodecimcostatus</Emphasis>)

In the present study, we analyzed the coexistence pattern of the Lusitanian pine vole (Microtus lusitanicus) and the Mediterranean pine vole (Microtus duodecimcostatus) in a potential area of sympatry in a Mediterranean landscape (Portugal). We also determined the relative contribution of local, landscape, and spatial factors explaining the differences in the distribution patterns of the two species in the region. Using a kriging interpolation method, we obtained a map of sympatric and allopatric areas of species occurrence. The estimated sympatry area corresponded to a northwest–southeast belt representing 11.3% of the study area. Habitat niche differences were assessed with binomial GLMs followed by a variance partitioning. At a local scale, higher altitude, higher cover of shrubs, lower clay content in the soil, and lower cover of tree canopy were the most important factors distinguishing M. lusitanicus presence sites from those with M. duodecimcostatus. At a larger scale, the presence of forest landscape units and the low abundance of “montado” units were the most influencing landscape factors in the identification of M. lusitanicus occurrence sites when compared to M. duodecimcostatus. Our results suggested that local coexistence of M. lusitanicus and M. duodecimcostatus in the field is a rare event. The differences in distribution patterns of the two pine vole species were mostly explained by fine-scale environmental factors and by shared spatial effects.     

Care of young, aggressiveness, and secretion of testosterone in male rodents: A correlation analysis

To test the current hypotheses on the relationship between the mating system, reproductive strategy, aggression, and secretion of testosterone, a comparative study of interactions in pair encounters, the level of parental care, and the gonadal testosterone level in males was performed in six rodent species (Clethrionomys rutilus, Meriones meridianus, Microtus arvalis, Lagurus lagurus, Lasiopodomys mandarinus, and Meriones unguiculatus) with different types of spatial-and-ethological population structures (SEPSs). It is shown that this dependence is absent in species with promiscuous mating and dominance hierarchy among males (C. rutilus and M. meridianus, SEPS type II). A trade-off, or negative correlation, was found in M. arvalis—a species with weak pair bonds and male competition for receptive females (SEPS type III). In species with persistent pair bonds and structured family groups (L. mandarinus and M. unguiculatus, SEPS type IV), no inverse relationship between the secretion of testosterone and paternal behavior was found either. Moreover, in male L. mandarinus androgens appear to stimulate paternal care.     

Organization and chromosomal localization of a B1-like containing repeat of Microtus subarvalis

A repetitive DNA sequence, MS2, was isolated from EcoRI-digested genomic DNA of the vole, Microtus subarvalis. The fragment was cloned and sequenced. Sequence analysis of this 1194-bp fragment revealed a 156-bp region demonstrating a 55% homology with the mouse B1 repeat. The remaining MS2 sequence shows no significant homology with other known GenBank sequences. The results of in situ hybridization of MS2 on vole metaphase chromosomes indicate the fragment is confined to heterochromatin blocks of the sex chromosomes in all but one species (M. arvalis). Distribution of MS2 sequences provides evidence for heterogeneity of the giant heterochromatin blocks of the XY Chromosomes (Chrs) in voles, for the unique cluster-like localization of MS2 within these blocks. Received: 10 October 1995 / Accepted: 30 March 1996     

Molecular and morphological evidence for multiple species within Paranoplocephala omphalodes (Cestoda, Anoplocephalidae) in Microtus voles (Arvicolinae)

Haukisalmi, V., Wickström, L. M., Henttonen, H., Hantula, J. & Gubányi, A. (2004). Molecular and morphological evidence for multiple species within Paranoplocephala omphalodes (Cestoda, Anoplocephalidae) in Microtus voles (Arvicolinae). —Zoologica Scripta, 33, 277–290. The present study was designed to test the hypothesis that the anoplocephalid cestode Paranoplocephala omphalodes (Hermann, 1783), a Holarctic parasite of Microtus voles, is a complex of host‐specific species, rather than a single host‐generalist species, using uni‐ and multivariate morphometrics and DNA sequence data from the mitochondrial cytochrome oxidase I gene. The phylogenetic methods applied to the mtDNA sequence data showed consistently that the cestodes morphologically recognizable as P. omphalodes include four well‐supported monophyletic groups, representing at least three distinct, largely host‐specific species. Multivariate morphometrics (discriminant analysis) successfully distinguished the four main mtDNA clades of P. omphalodes‐like cestodes. The true P. omphalodes is shown to be a parasite of Microtus arvalis, M. agrestis and Clethrionomys glareolus in Europe. Microtus oeconomus harbours two host‐specific, allopatric and possibly conspecific clades, one with a Holarctic and another with an (eastern) Beringian (Alaskan) distribution. The eastern Beringian endemic M. miurus is also parasitized with a host‐specific, morphologically divergent species of Paranoplocephala. The cestode clades recognized in M. oeconomus and M. miurus represent 2–3 undescribed species. Molecular phylogenetic analyses supported the monophyly of the ‘northern clade’ of Paranoplocephala spp., an assemblage including P. kalelai from Clethrionomys spp., P. macrocephala from Microtus spp. and all clades of P. omphalodes‐like cestodes except those representing the true P. omphalodes from Europe. The intra‐ and interspecific phylogeny within the northern clade is compared tentatively with the known evolutionary history of the hosts.     

Demography of a mediterranean microtine: the Mediterranean pine vole,Microtus duodecimcostatus

Microtus duodecimcostatus in a mediterranean vole which is not known to display spectacular increases in population numbers as in some microtine species. A population was studied in southern France with a capture-recapture method. The population included resident adults which have a high and constant survival rate (monthly estimate: 0.879), erratic adults (those caught once only), and juveniles which have a lower and constant survival rate. The adult survival rate was not sexbiased but the juvenile survival rate was higher in males (monthly estimates: 0.710 and 0.596 for males and females, respectively). Adult body weight did not vary seasonally. Residents had a higher mean body weight than erratics. Reproduction occurred all the year round. The proportion of reproductive females was higher among residents than among erratics. Population numbers varied seasonally. Our study points out thatM. duodecimcostatus is very different from microtine species which display cyclic fluctuations. Population studies on the subgenusPitymys (which containsM. duodecimcostatus and its closest related species) suggest that they are typically non-cyclic. The importance of social factors in the control of reproduction and maturation was evidenced inM. pinetorum. The role of such factors in the population regulation ofM. duodecimcostatus is discussed.     

Über die seitendrüsen der mitteleuropäischen wühlmäuse der gattung Microtus Schrank

Zusammenfassung Es werden bei drei Wühlmausarten Hautdrüsenorgane am hinteren Rücken beschrieben, bei Microtus agrestis und Pitymys subterraneus erstmalig. In der Ausprägung stimmen die Strukturen bei den drei Arten nicht völlig überein, obwohl es sich bei den tragenden Elementen in allen Fallen um hypertrophische und abgewandelte Haarbalgtalgdrüsen handelt. Über die Funktion dieser Steißdrüsen liegen keine Beobachtungen vor. Bei der Feldmaus (Microtus arvalis) konnten bisher keine Hinweise auf Hautdrüsenorgane gefunden werden.Vortrag, gehalten auf der 39. Hauptversammlung der Deutschen Gesellschaft für Säugetierkunde in Braunschweig, am 6. 10. 65.     

Genomic versus chromosomal polytypy in studies of mitochondrial and nuclear DNA markers in the <Emphasis Type="Italic">Microtus arvalis</Emphasis> group

Common voles of the Microtus arvalis group distributed over the territory of European Russia are represented by three karyotypic categories, i.e., sympatric sibling species with 2n = 46 and 54, and two allopatric karyoforms in one of them, 2n = 46. For each category, molecular markers were found. For two 46-chromosome forms (arvalis and obscurus), iDNA was for the first time studied in karyotyped and non-karyotyped specimens for a parapatric hybrid zone, where high diversity of intermediate karyotypes was recorded. Preferential migration of the mitochondrial markers in arvalis and significant differences in the cline width for chromosomal and nuclear markers in obscurus were shown. The hybrid zone examined exhibited unusual combination of such features as the practically complete absence of “pure” representatives of the original parental forms and a clear deficiency of the first generation hybrids. The mtDNA divergence for the arvalis and obscurus karyoforms (4.6%) is comparable to the lowest limit for interspecies differences within the large and complex genus Microtus.     

Evidence for multiple functional copies of the male sex-determining locus, sry, in African murine rodents

Southern hybridization data suggest that the male sex-determining locus, Sry, is often duplicated in rodents. Here we explore DNA sequence evolution of orthologous and paralogous copies of Sry isolated from six species of African murines. PCR amplification followed by direct sequencing revealed from two to four copies of Sry per species. All copies include a long open reading frame, with a stop codon that coincides closely with the stop codon of the house mouse, Mus musculus, a species known to have a single copy of Sry. A phylogenetic analysis suggests that there are at least seven paralogous copies of Sry in this group of rodents. Putative orthologues are identical; sequence divergence among putative paralogues ranges from 1 to 8% (excluding the CAG repeat), with much lower levels of divergence in the high-mobility group (HMG-box) region than in the C-terminal region. A high proportion of nucleotide substitutions in both regions result in amino-acid replacement. The long open reading frame, conserved HMG-box, and pattern of evolution of the putative paralogues suggest that they are functional. Received: 4 October 1996 / Accepted: 17 January 1997     

Sry‐negative XX sex reversal in purebred dogs

The gene responsible for testis induction in normal male mammals is the Y‐linked Sry. However, there is increasing evidence that other genes may have testis‐determining properties. In XX sex reversal (XXSR), testis tissue develops in the absence of the Y chromosome. Previous polymerase chain reaction (PCR) assays indicated that autosomal recessive XXSR in the American cocker spaniel is Sry‐negative. In this study, genomic DNA from the breeding colony of American cocker spaniels and from privately owned purebred dogs were tested by PCR using canine primers for the Sry HMG box and by Southern blots probed with the complete canine Sry coding sequence. Sry was not detected by either method in genomic DNA of affected American cocker spaniels or in the majority (20/21) of affected privately owned purebred dogs. These results confirm that the autosomal recessive form of XXSR in the American cocker spaniel is Sry‐negative. In combination with previous studies, this indicates that Sry‐negative XXSR occurs in at least 15 dog breeds. The canine disorder may be genetically heterogeneous, potentially with a different mutation in each breed, and may provide several models for human Sry‐negative XXSR. A comparative approach to sex determination should be informative in defining the genetic and cellular mechanisms that are common to all mammals. Mol. Reprod. Dev. 53:266–273, 1999. © 1999 Wiley‐Liss, Inc.     

HMG-box sequences from microbats homologous to the human SOX30 HMG-box*

SOX genes are a family of genes that encode for proteins which are characterised by the presence of a HMG-domain related to that of the mammalian sex-determining gene (SRY). By definition, the DNA binding domain of SOX genes is at least 50% identical to the 79 amino acid HMG domain of the SRY gene. We report here two HMG-box sequences from two microbat species (R. ferrumequinum and P. Pipistrellus) which were PCR amplified using a primer pair specific to the mouse Sry HMG-box. The high percentage of identity of this sequences with the human and mouse SOX30 HMG-box suggests that they are the SOX30 HMG-box for these two bat species. The sequencing data reported in this paper are available in the EMBL Nucleotide Sequence Database through the following accession numbers: Pipistrellus pipistrellus SOX30 gene: AJ243292; Rhinolophus ferrumequinum SOX30 gene: AJ243293. This revised version was published online in July 2006 with corrections to the Cover Date.     

Isolation of ES-Like Lines of Common Voles of the Genus Microtus from Blastocysts and Germ Cells and as a Result of Fusion of Somatic Cells with Mouse Embryonic Stem Cells

Three and four independent cell lines with limited pluripotency were obtained from the inner cell mass cells of blastocysts and primordial germ cells of common voles, respectively. The results of cytogenetic analysis suggest that all these lines originated from the embryos of F₁ Microtus rossiaemeridionalis × M. arvalis males and had a great number of near-triploid cells already during the early passages. The cells of these lines, like those of the inner cell mass, were characterized by the alkaline phosphatase activity. Nine independent cell lines were obtained as a result of hybridization of the mouse embryonic stem cells and vole splenocytes: eight lines and one line from hybridization with the M. kirgisorum and M. rossiaemeridionalis splenocytes, respectively. The cells of these lines expressed some properties of embryonic stem lines had a chromosome complement similar to the sum of two initial diploid sets of the mouse and vole.     

Somatic characteristics and reproduction of common vole, <Emphasis Type="Italic">Microtus arvalis</Emphasis> (Microtus arvalis

Reproduction potential and biometry of somatic characteristics of the common vole Microtus arvalis were evaluated and discussed. The results were processed on the basis of an extensive material (2,171 individuals) from the whole territory of Slovakia (315 sites situated at altitudes from 100 to 1500 m above sea level). Among the somatic characteristics studied, the highest variability was found in body length and the smallest in hind foot length. Highly significant differences were also found between the foot length of adult males and females. Populations of M. arvalis at low altitudes were less numerous than at higher altitudes. Altitudinal differences in average embryo numbers in female uteri as well as differences in follicle length in males during the reproductive season were also observed.     

Genes for the SMC Family Proteins in a Common Vole Microtus arvalis

Genes for four subfamilies of SMC (structural maintenance of chromosomes) proteins have been isolated from the genome of a common vole Microtus arvalis. The high degree of homology between representatives of each SMC protein subfamily of different classes of organisms has been demonstrated. The full-sized copy of a mammalian gene encoding SMC4 protein has been isolated and analyzed for the first time. The SMC proteins enter into the composition of complexes responsible for cohesion of sister chromatids, formation of mitotic chromosomes, recombination, DNA repair, and regulation of gene expression. We discuss the possible participation of the SMC proteins in inactivation of the X chromosome in mammalian females. Common voles of genus Microtusgroup arvalis serve a unique model for the study of the inactivation process.     

Sexing and detection of gene construct in microinjected bovine blastocysts using the polymerase chain reaction

We present a polymerase chain reaction (PCR)-based procedure for rapid bovine embryo sexing and classifying embryos for the presence of exogenous DNA. Fourteen bovine blastocysts microinjected with gene construct DNA at the pronuclear stage were divided into quarters and subjected to amplification with construct-specific and sex gene-specific (ZFY/ZFX) primers in the same initial PCR reaction. Blastocysts carrying microinjected construct DNA could be identified by the presence of construct-specific PCR product in approximately 4 h. Approximately half of the microinjected and two of 16 non-microinjected blastocysts typed PCR-positive for the construct DNA. Owing to erroneous amplifications in the two non-microinjected control blastocysts, and the inability of the system to distinguish integrated from non-integrated copies of the microinjected construct, the number of construct-positive blastocysts determined in our assay most likely overestimates the number of true transgenic embryos. Nevertheless, using this assay, we were able to determine that approximately half of the microinjected embryos were negative for the transgene construct and thus could be eliminated from transfer to a recipient cow. Embryo sexing was achieved in less than 6 h by restriction fragment length polymorphism analysis of nestedZFY/ZFXPCR products reamplified from initial PCR reactions. In 11/14 microinjected blastocysts all sections assayed unambiguously as the same sex. In one embryo, only one section was analysed, while two other blastocysts whowed some discrepancies of sexing results between the sections analysed. The approach employed here to determine the sex and presence of microinjected construct DNA in bovine preimplantation embryos is rapid, accurate among different sections of an embryo and can be used to increase the efficiency of current transgenic cattle production procedures.     

Isolation and characterization of polymorphic microsatellite loci in the field vole,Microtus agrestis,and their cross‐utility in the common vole,Microtus arvalis

We developed nine polymorphic microsatellite loci for the field vole, Microtus agrestis. The number of alleles ranged from five to 15 and observed heterozygosities ranged from 0.40 to 1.00. We also tested the microsatellite loci for amplification and polymorphism in the congeneric species Microtus arvalis. Five of the nine loci were successfully analysed in this species. The microsatellite markers will be employed in studies of reproductive success and fine‐scale spatial genetic structure.